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A rapid inoculation and identification method for citrus canker

A technology for citrus canker and identification method, which is applied in the field of rapid inoculation and identification of citrus canker, can solve the problems of insufficient nutrient and water supply of leaves, complicated management of water, fertilizer, pests and diseases, and large space for cultivation environment, and achieves rapid and efficient screening, Reliable identification results and the effect of reducing the influence of environmental factors

Active Publication Date: 2022-04-01
HUNAN AGRICULTURAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Both in vitro and in vivo identification methods have different defects. For example, in vitro identification means that the leaves are separated from the mother tree, the supply of nutrients and water to the leaves is insufficient, the disease is unstable, and the leaves are easy to dry and die; although the in vivo identification is relatively stable, the operation Inconvenient, the required cultivation environment takes up a lot of space, the demand for inoculated seedlings is also large, and the management of water, fertilizer, diseases and insect pests is more complicated

Method used

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  • A rapid inoculation and identification method for citrus canker
  • A rapid inoculation and identification method for citrus canker
  • A rapid inoculation and identification method for citrus canker

Examples

Experimental program
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Effect test

Embodiment 1

[0027] The operation steps of this embodiment are as follows:

[0028] (1) Sample collection: Pick 3 relatively tender leaves that have reached the maximum leaf area but have not completely turned green from the shoots of the plants, put them in a ziplock bag and mark the samples; -05 and the susceptible variety Bingtang Orange were used as controls;

[0029] (2) Pathogenic bacteria culture: Pick a single colony of ulcer disease from the preserved solid medium plate, add 20 mL liquid LB medium, shake and cultivate in a constant temperature shaker at 28°C, 200 r / min for 20 h, use sterile water Dilute the pathogen suspension to 10 3 、10 4 、10 5 、10 6 cfu / ml;

[0030] (3) Identification of canker disease by sample inoculation: After the sample is taken back to the laboratory, wipe the front and back of the leaf with alcohol cotton, turn the leaf so that the back of the leaf is facing upward, and after the alcohol evaporates, use a 1 mL syringe needle in the middle of the vei...

Embodiment 2

[0035] (1) Sample collection: Pick 3 relatively tender leaves that have reached the maximum leaf area but have not completely turned green from the shoots of the plants, put them in a ziplock bag and mark the samples; -05 and the susceptible variety Bingtang Orange were used as controls;

[0036] (2) Pathogenic bacteria culture: Pick a single colony of ulcer disease from the preserved solid medium plate, add 20 mL liquid LB medium, shake and cultivate in a constant temperature shaker at 28°C, 200 r / min for 20 h, use sterile water Dilute the pathogen suspension to 10 3 、10 4 、10 5 、10 6 cfu / ml;

[0037] (3) Identification of canker disease by sample inoculation: After the sample is taken back to the laboratory, wipe the front and back of the leaf with alcohol cotton, turn the leaf so that the back of the leaf is facing upward, and after the alcohol evaporates, use a 1 mL syringe needle in the middle of the veins on both sides. Puncture a small hole (do not pierce the leaf)...

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Abstract

A method for rapid inoculation and identification of citrus canker disease, comprising the following steps: (1) sample leaf collection; (2) cultivation of citrus canker pathogenic bacteria; (3) sample leaf inoculation and identification of canker disease; (4) sample leaf rooting culture; 5) Sample leaf culture management. Aiming at the shortcomings and deficiencies of the current citrus canker resistance identification, the invention innovates the cultivation and identification method by in vitro culturing and inoculating the leaves of canker to take root, supplementing nutrients and water to increase the growth and observation cycle. The invention has the advantages of simple operation, efficient and rapid inoculation and identification, reliable results and more intuitive.

Description

technical field [0001] The invention relates to a detection method for citrus canker, in particular to a rapid inoculation and identification method for citrus canker. Background technique [0002] Citrus is the largest fruit in the world, and citrus canker is a worldwide quarantine disease that seriously hinders the development of citrus industry, and its prevention and control is difficult. [0003] Citrus canker is caused by Xanthomonas citri subsp. citri, its growth temperature is 5~36℃, the most suitable temperature for the growth of canker bacteria is 25~30℃, and the lethal temperature is 55~65℃ ℃. Because Citrus bacterial canker disease is difficult to control and cannot be eradicated, and the pathogen has a strong ability to spread, once it occurs in a new area, it will require a lot of manpower and material resources to prevent and control, causing great economic losses. Breeding disease-resistant varieties is the only effective way to prevent and control canker. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/18A01H4/00C12N1/20C12R1/64
CPCA01H4/008C12N1/20C12Q1/18
Inventor 马先锋朱志媚邓子牛戴素明盛玲
Owner HUNAN AGRICULTURAL UNIV
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